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Bradford Protein Concentration Assay

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摘要:BradfordProteinConcentrationAssayversion01/07/2001Abbreviations:mcg=microgramsmcL=microlitersBSA=bovineserumalbuminO。=opticaldensitydI=deionizedBackgroundTheBradfordproteinassay(1)isoneofseveralsimplemethodscommonlyusedtodeterminethetotalprot......

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Bradford Protein Concentration Assay
version 01/07/2001

Abbreviations:

Background

The Bradford protein assay (1) is one of several simple methods commonly used to determine the total protein concentration of a sample. The method is based on the proportional binding of the dye Coomassie to proteins. Within the linear range of the assay (~5-25 mcg/mL), the more protein present, the more Coomassie binds. Furthermore, the assay is colorimetric; as the protein concentration increases, the color of the test sample becomes darker. Coomassie absorbs at 595 nm. The protein concentration of a test sample is determined by comparison to that of a series of protein standards known to reproducibly exhibit a linear absorbance profile in this assay. Although different protein standards can be used, we have chosen the most widely used protein as our standard - Bovine Serum Albumin (BSA).

Procedure

Table 1. Preparation of test samples for the Bradford protein assay.

Test Sample
Sample Volume,
mcL
Vol. Water,
mcL
Vol. Bradford Reagent,
mcL
Reference Cell
0
800
200
Blank
0
800
200
BSA Standard - 5 mcg/mL
10
790
200
BSA Standard - 10 mcg/mL
20
780
200
BSA Standard - 15 mcg/mL
30
770
200
BSA Standard - 20 mcg/mL
40
760
200
BSA Standard - 25 mcg/mL
50
750
200
Protein Sample
50
750
200

作者: 佚名 2008-2-3
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